Crossbridges in insect flight muscles of the blowfly (Sarcophaga bullata)

Abstract

Crossbridges in quick-frozen deep-etched blowfly flight muscles (from Sarcophaga bullata) were compared with those observed in the traditional waterbug preparation (Lethocerus) and found to be indistinguishable. Hence, blowfly was chosen as a fresher more accessible tissue for determining the effect of various fixatives and nucleotides on crossbridge structure. In rigor control, crossbridges were most regular in muscles that were stabilized before freezing by prefixation in glutaraldehyde followed by 'hardening' with neutralized tannic acid, so all nucleotide treatments were terminated by such fixation. MgATP (5 mM) converted the rigor pattern of crossbridges into a random array of disconnected thick filament projections. Lower levels of ATP (0.1 mM) caused a variable but generally lesser degree of crossbridge disconnection, as did 5 mM ADP (probably because it slowly converted to ATP inside the muscle fibres). Vanadate (1-2 mM) potentiated muscle relaxation in the latter two nucleotide treatments (i.e. produced a greater degree of crossbridge disconnection). Thus, differences in overall crossbridge abundance were readily apparent in chemically fixed muscles. Structural details within individual crossbridges were less well preserved, however. Chemical prefixation tended to collapse the muscle lattice, add a surface film to the filaments and thus obscure crossbridge details. Rigorous control of fixative pH largely prevented these problems and permitted recognition of the fact that in Sarcophaga flight muscle, as in Lethocerus muscle in rigor, the S1 'heads' of crossbridges attach to the thin filaments in the expected 'arrowhead' configuration.